Understanding abiotic stress responses is one of the most important issues in plant research nowadays. Abiotic stress, including excess light, can promote the onset of oxidative stress through the accumulation of reac...Understanding abiotic stress responses is one of the most important issues in plant research nowadays. Abiotic stress, including excess light, can promote the onset of oxidative stress through the accumulation of reactive oxygen species. Oxidative stress also arises when in vitro propagated plants are exposed to high light upon transfer to ex vitro. To determine whether the underlying pathways activated at the transfer of in vitro grapevine to ex vitro conditions reflect the processes occurring upon light stress, we used Vitis vinifera Affymetrix GeneChip （VvGA） and a custom array of genes responsive to light stress （LSCA） detected by real-time reverse transcriptase PCR （qRT-PCR）. When gene-expression profiles were compared, ＇protein metabolism and modification＇, ＇signaling＇, and ＇anti-oxidative＂ genes were more represented in LSCA, while, in VvGA, ＇cell wall metabolism＇ and ＇secondary metabolism＇ were the categories in which gene expression varied more significantly. The above functional categories confirm previous studies involving other types of abiotic stresses, enhancing the common attributes of abiotic stress defense pathways. The LSCA analysis of our experimental system detected strong response of heat shock genes, particularly the protein rescuing mechanism involving the cooperation of two ATP-dependent chaperone systems, Hsp100 and Hsp70, which showed an unusually late response during the recovery period, of extreme relevance to remove non-functional, potentially harmful polypeptides arising from misfolding, denaturation, or aggregation brought about by stress. The success of LSCA also proves the feasibility of a custommade qRT-PCR approach, particularly for species for which no GeneChip is available and for researchers dealing with a specific and focused problem.展开更多
目的:揭示肝癌诱发过程中肝组织基因表达谱的演变,为中医药防治原发性肝癌提供参考。方法:采用DEN诱发大鼠肝癌,分别于诱癌的第4周、8周、16周、20周(肝癌形成)切取肝(合肝癌)组织,常规提取RNA,Affymetrix Rat 230A GeneChip及技术检测...目的:揭示肝癌诱发过程中肝组织基因表达谱的演变,为中医药防治原发性肝癌提供参考。方法:采用DEN诱发大鼠肝癌,分别于诱癌的第4周、8周、16周、20周(肝癌形成)切取肝(合肝癌)组织,常规提取RNA,Affymetrix Rat 230A GeneChip及技术检测大鼠肝组织基因表达的差异和演变。结果:在芯片的15710个基因中,正常组有9225个基因表达,诱癌4周表达增至9396个,8周增至9872个,16周增至10496个,20周有10420个。在肝癌诱发过程中,存在大量基因表达的消长以及高表达基因数的增加,其中部分已知基因的结果。结论:DEN诱发大鼠肝癌过程中基因组变化是十分复杂的,而且文献追踪表明,国内外对在这些基因的功能及其与肝癌发生、发展的关系大多不明确。因此,如何逐一找到那些起着关键作用的基因,进一步阐释其在肝癌的发生、发展和转归中的作用,及其与中医证候演变和相应治法的关系,是今后中医基础理论实验研究的重要方向。展开更多
文摘Understanding abiotic stress responses is one of the most important issues in plant research nowadays. Abiotic stress, including excess light, can promote the onset of oxidative stress through the accumulation of reactive oxygen species. Oxidative stress also arises when in vitro propagated plants are exposed to high light upon transfer to ex vitro. To determine whether the underlying pathways activated at the transfer of in vitro grapevine to ex vitro conditions reflect the processes occurring upon light stress, we used Vitis vinifera Affymetrix GeneChip （VvGA） and a custom array of genes responsive to light stress （LSCA） detected by real-time reverse transcriptase PCR （qRT-PCR）. When gene-expression profiles were compared, ＇protein metabolism and modification＇, ＇signaling＇, and ＇anti-oxidative＂ genes were more represented in LSCA, while, in VvGA, ＇cell wall metabolism＇ and ＇secondary metabolism＇ were the categories in which gene expression varied more significantly. The above functional categories confirm previous studies involving other types of abiotic stresses, enhancing the common attributes of abiotic stress defense pathways. The LSCA analysis of our experimental system detected strong response of heat shock genes, particularly the protein rescuing mechanism involving the cooperation of two ATP-dependent chaperone systems, Hsp100 and Hsp70, which showed an unusually late response during the recovery period, of extreme relevance to remove non-functional, potentially harmful polypeptides arising from misfolding, denaturation, or aggregation brought about by stress. The success of LSCA also proves the feasibility of a custommade qRT-PCR approach, particularly for species for which no GeneChip is available and for researchers dealing with a specific and focused problem.
文摘目的:揭示肝癌诱发过程中肝组织基因表达谱的演变,为中医药防治原发性肝癌提供参考。方法:采用DEN诱发大鼠肝癌,分别于诱癌的第4周、8周、16周、20周(肝癌形成)切取肝(合肝癌)组织,常规提取RNA,Affymetrix Rat 230A GeneChip及技术检测大鼠肝组织基因表达的差异和演变。结果:在芯片的15710个基因中,正常组有9225个基因表达,诱癌4周表达增至9396个,8周增至9872个,16周增至10496个,20周有10420个。在肝癌诱发过程中,存在大量基因表达的消长以及高表达基因数的增加,其中部分已知基因的结果。结论:DEN诱发大鼠肝癌过程中基因组变化是十分复杂的,而且文献追踪表明,国内外对在这些基因的功能及其与肝癌发生、发展的关系大多不明确。因此,如何逐一找到那些起着关键作用的基因,进一步阐释其在肝癌的发生、发展和转归中的作用,及其与中医证候演变和相应治法的关系,是今后中医基础理论实验研究的重要方向。
